Generation of T-cell receptor retrogenic mice
Generation of T-cell receptor retrogenic mice
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DOI：
10.1038/nprot.2006.61
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发表时间：
2006-01-01
期刊：
影响因子：
14.8
通讯作者：
Vignali, Dario A. A.
中科院分区：
文献类型：
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作者：
Holst, Jeff;Szymczak-Workman, Andrea L.;Vignali, Dario A. A.
T细胞受体(TCR)转基因(TG)小鼠彻底改变了我们对T细胞生物学许多方面的认识。尽管它们提供了几乎无限的单一特异性T细胞来源，但将它们培育到不同的背景和/或新的基因敲除/敲入小鼠模型通常是耗时的(6个月到几年)，这可能会使这一过程成本高昂，并可能显著推迟研究。该方案描述了一种新的方法，利用逆转录病毒介导的干细胞基因转移，从单个2A多肽连接的多顺反子逆转录病毒载体中表达定义的TCR-α和TCR-β蛋白。为了避免与传统转基因小鼠混淆，我们将这些小鼠称为‘逆转录’(Rg)小鼠(‘retro’来自逆转录病毒，‘Genetic’来自TG)。我们已经成功地使用这种方法在短短6周内在几种不同的小鼠背景上表达了50多个不同的TCR。
T-cell receptor (TCR) transgenic (Tg) mice have revolutionized our understanding of many aspects of T-cell biology. Whereas they provide an almost unlimited source of T cells with a single specificity, breeding them onto different backgrounds and/or new knockout/knock-in mouse models is often time-consuming ( 6 months to several years), which can make the process costly and can significantly delay research. This protocol describes a new method for expressing defined TCR-alpha and TCR-beta proteins from a single 2A peptide-linked multicistronic retroviral vector in mice, using retrovirus-mediated stem cell gene transfer. We refer to these as 'retrogenic' (Rg) mice ('retro' from retrovirus and 'genic' from Tg) to avoid confusion with traditional transgenic mice. We have successfully used this approach to express over 50 different TCRs on several different mouse backgrounds in as little as 6 weeks.